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PMID: 8491203 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Membrane glycoprotein folding, oligomerization and intracellular transport: effects of dithiothreitol in living cells.

The EMBO journal ·Vol. 12 ·No. 5 ·1993-05-00 ·Pages 2151-7

Tatu U, Braakman I, Helenius A

Abstract

Using influenza hemagglutinin (HA0) and vesicular stomatitis virus G protein as model proteins, we have analyzed the effects of dithiothreitol (DTT) on conformational maturation and transport of glycoproteins in the secretory pathway of living cells. While DTT caused reduction of folding intermediates and misfolded proteins in the endoplasmic reticulum (ER), it did not affect molecules that had already acquired a mature trimeric conformation, whether present in the ER or elsewhere. The conversion to DTT resistance was therefore a pre-Golgi event. Reduction of folding intermediates was dependent on the intactness of the ER and on metabolic energy, suggesting cooperativity between DTT and ER folding factors. DTT did not inhibit most cellular functions, including ATP synthesis and protein transport within the secretory pathway. The results established DTT as an effective tool for analyzing the folding and compartmental distribution of proteins with disulfide bonds.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Biological Transport/drug effects CHO Cells Cell Membrane/drug effects,metabolism Cricetinae Dithiothreitol/pharmacology Endoplasmic Reticulum/drug effects,metabolism Golgi Apparatus/drug effects,metabolism Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral/chemistry,metabolism Membrane Glycoproteins/chemistry,metabolism Protein Folding Viral Envelope Proteins/chemistry,metabolism
Chemicals
G protein, vesicular stomatitis virus Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral Membrane Glycoproteins Viral Envelope Proteins Adenosine Triphosphate Dithiothreitol
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tatu U
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510.
Braakman I
Helenius A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-05-00
Pages
2151-7
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413436
Subset
IM
Grants
NCI NIH HHS · 5P01 CA 46182 · United States
NIGMS NIH HHS · 5R01 GM 38346 · United States
External Links
PubMed source PMC Full Text View journal details
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